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作物学报 ›› 2018, Vol. 44 ›› Issue (6): 796-813.doi: 10.3724/SP.J.1006.2018.00796

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

玉米雌穗发育杂种优势相关miRNA的研究

王琪月1,孟淑君1,张柯2,张战辉1,汤继华1,丁冬1,*()   

  1. 1 河南农业大学农学院 / 省部共建小麦玉米作物学国家重点实验室, 河南郑州 450002
    2 河南省农业科学院, 河南郑州450002
  • 收稿日期:2017-09-29 接受日期:2018-03-26 出版日期:2018-06-12 网络出版日期:2018-04-18
  • 通讯作者: 丁冬
  • 基金资助:
    本研究由国家自然科学基金项目(31370033);国家高技术研究发展计划项目资助(2012AA10A305)

Investigation of Maize miRNA Involved in Developing-ear Heterosis

Qi-Yue WANG1,Shu-Jun MENG1,Ke ZHANG2,Zhan-Hui ZHANG1,Ji-Hua TANG1,Dong DING1,*()   

  1. 1 College of Agronomy, Henan Agricultural University / Key Laboratory of Wheat and Maize Crops Science, Zhengzhou 450002, Henan, China
    2 Henan Academy of Agricultural Sciences, Zhengzhou 450002, Henan, China
  • Received:2017-09-29 Accepted:2018-03-26 Published:2018-06-12 Published online:2018-04-18
  • Contact: Dong DING
  • Supported by:
    This study was supported by the National Natural Science Foundation of China(31370033);the National High Technology Research and Development Program of China(2012AA10A305)

摘要:

杂种优势已广泛应用于玉米育种, 在提高玉米产量、品质以及增强抗逆性等方面起到了重要的作用, 然而杂种优势的分子机制尚不清楚。植物miRNA主要在转录和转录后水平调节基因的表达。为阐明miRNA是否及如何对玉米雌穗发育杂种优势产生影响, 本研究对玉米杂交种郑单958及其亲本自交系(昌7-2和郑58)进行了高通量miRNA测序和降解组测序。取玉米雌穗花序分生组织(IM)发育为成对小穗分生组织(SPM), 进而产生小穗分生组织(SM), 及小花分生组织(FM)将3个不同时期的雌穗样品用于miRNA建库测序, 鉴定出16个miRNA家族中的81个保守miRNA为非加性表达, 认为是与雌穗发育杂种优势形成相关的miRNA; 3个阶段中分别检测到80.30%、56.06%和48.10%的非加性表达的miRNA被显性或超显性抑制。鉴定出8种新的miRNA, 属于7个miRNA家族。通过雌穗降解组测序, 发现在郑单958及其亲本自交系中共同检测到的miRNA靶向42个基因的82个转录本。根据测序结果构建了miRNA参与玉米雌穗杂种优势的模型, 并推测在雌穗发育早期阶段杂交种雌穗的miRNA的普遍抑制导致其靶基因上调表达, 随着发育进程miRNA逐步解除抑制, 带来其靶基因表达量的逐步减少, 这种miRNA与其靶基因的拮抗关系也许与玉米雌穗发育杂种优势形成有关。

关键词: 雌穗发育, 杂种优势, miRNA, 降解组测序, 玉米

Abstract:

Heterosis has been utilized widely in maize breeding; it plays an important role in increasing yield, improving quality and enhancing stresses resistance. However, the molecular mechanism of heterosis is far from clear. MicroRNAs (miRNAs) act as key regulating factors of gene expression in post-transcriptional level. To investigate whether miRNA-dependent gene regulation is responsible for heterosis during maize ear development, we performed an illumine miRNA deep-sequencing on the most widely-planted elite hybrid Zhengdan 958 and its parental inbred lines (Chang 7-2 and Zheng 58) in China. The ear samples at the developmental stages of inflorescence meristem (IM) producing spikelet pair meristems (SPM), resulting in spikelet meristems (SMs), and floral meristems (FM) were collected for RNA library construction. As a result, 81 conserved miRNAs belonging to 16 miRNA families were identified which were non-additively expressed. At the three stages, 80.30%, 56.06%, and 48.10% of these non-additively expressed miRNAs were repressed ever dominantly or over-dominantly. A total of 82 target transcripts from 42 genes of conserved miRNAs among Zhengdan 958 and its parents were detected via whole genome degradome sequencing. Additionally, eight novel maize miRNAs belonging to seven miRNA families were identified using stringent criteria. Global repression of miRNAs in hybrid ears at the early stage might lead to the up-regulated expressing of their target genes. Moreover, the in-step de-repression of given miRNA family members may be the reason of enhanced repression of their target genes. These results revealed, at least in part, the involvement of maize miRNAs in hybrids leads to higher ear developing vigor compared with its parental lines.

Key words: ear development, heterosis, miRNA, degradome, Zea mays L.

表1

玉米雌穗杂种优势相关miRNA和其靶基因的检测引物序列"

引物
Primer
引物序列
Primer sequence (5°-3°)
zma-miR159a/b RT F: GTCGTATCCAGTGCAGGGTCCGAGG; R: TATTCGCACTGGATACGACCAGAGC
zma-miR159a/b PCR CGACCTTTGGATTGAAGGGA
zma-miR160a-e,gRT F: GTCGTATCCAGTGCAGGGTCCGAGG; R: TATTCGCACTGGATACGACTGGCAT
zma-miR160a-e,gPCR GTAAGTGCCTGGCTCCCTGT
zma-miR167a-dRT F: GTCGTATCCAGTGCAGGGTCCGAGG; R: TATTCGCACTGGATACGACTAGATC
zma-miR167a-dPCR TCTAGTGAAGCTGCCAGCAT
zma-miR168a/b RT F: GTCGTATCCAGTGCAGGGTCCGAGG; R: TATTCGCACTGGATACGACGTCCCG
zma-miR168a/b PCR CATAGTCGCTTGGTGCAGAT
zma-miR169a/b RT F: GTCGTATCCAGTGCAGGGTCCGAGG; R: TATTCGCACTGGATACGACTCGGCA
zma-miR169a/b PCR CGTAGCAGCCAAGGATGACT
zma-miR172a-dRT F: GTCGTATCCAGTGCAGGGTCCGAGG; R: TATTCGCACTGGATACGACTGCAGC
zma-miR172a-dPCR GCGCCAGAATCTTGATGAT
zma-miR172eRT F: GTCGTATCCAGTGCAGGGTCCGAGG; R: TATTCGCACTGGATACGACATGCAG
zma-miR172ePCR CCGTCGGAATCTTGATGATG
zma-miR390a/b RT F: GTCGTATCCAGTGCAGGGTCCGAGG; R: TATTCGCACTGGATACGACGGCGCT
zma-miR390a/b PCR GCATCAAGCTCAGGAGGGAT
Universal Reverse ATTCATGAAAGGCGGATGGC
Y-tubulin-A TGTCGTCCAACCTTACAACTCACT
Y-tubulin-S TCTCCAGGGTCCTCCATTCC
T159a/b(GRMZM2G423833_T01)-A TGGTCTGGCTGACTGTTGTT
T159a/b(GRMZM2G423833_T01)-S CTTGTGAGCGTGATGGAAAT
T160a-e,g(AC207656.3_FGT002)-A AACCGAAACTGCCGAACC
T160a-e,g(AC207656.3_FGT002)-S TGACCATACTGACGGACGAGA
T167a-d(GRMZM2G078274_T03)-A CAGGGATGGATACATAGGAAAG
T167a-d(GRMZM2G078274_T03)-S GAATCAACCGCAGGAGAAAG
#T168a/b(GRMZM2G361518_T01)-A ACAACAGCAGCAATGGAAGG
#T168a/b(GRMZM2G361518_T01)-S AAGGTCGGAGGAAGGAACAC
T169a/b(GRMZM2G000686_T01)-A TATGATATGGCGAAGGACTG
T169p(GRMZM2G000686_T01)-S CATCAAGAAGCGTCTGGAAT
T172a-d(GRMZM5G862109_T01)-A GCGATAAATAGCGTGGTGGG
T172a-d(GRMZM5G862109_T01)-S CTTCTCGTCCTCGGTCCTCC
T172e(GRMZM2G700665_T02)-A CGTTGTTGCGGTACATTTAGGC
T172e(GRMZM2G700665_T02)-S GTGCAGGATGGTCGTCAGGG
T390a/b(GRMZM2G084821_T02)-A CCCGGTGGCATAATAACTGC
T390a/b(GRMZM2G084821_T02)-S TGGAGCCGAATAATGGAAGC

图1

miRNA的长度分布"

表2

共检测到的保守miRNA的表达趋势"

miRNA IM to SPM SPM to SM FM miRNA IM to SPM SPM to SM FM
zma-miR156a-5p - - - - - zma-miR166m-3p - - + - + +
zma-miR156b-5p - - - - - - zma-miR166n-3p - - + - -
zma-miR156c - - - - - zma-miR167a-5p + - - - - -
zma-miR156d-5p - - - - - - zma-miR167b-5p + - - - - -
zma-miR156f-5p - - - - - - zma-miR167c-5p + - - - - -
zma-miR156g-5p - - - - - - zma-miR167d-5p + - - - - -
zma-miR156j-5p + + + - - zma-miR167e-5p + - - - - -
zma-miR156k-5p - - - - - - zma-miR167f-5p + - - - - -
zma-miR159a-3p - - - + + zma-miR167g-5p + - - - - -
zma-miR159b-3p - - - + + zma-miR167h-5p + - - - - -
zma-miR159f-3p - - - + + zma-miR167i-5p + - - - - -
zma-miR159j-3p - - - + + zma-miR167j-5p + - - - - -
zma-miR159k-3p - - - + + zma-miR168a-5p + - + + + +
zma-miR160a-5p - - + + + + zma-miR168b-5p + - + + + +
zma-miR160b-5p - - + + + + zma-miR169a-5p - - + + + +
zma-miR160c-5p - - + + + + zma-miR169b-5p - - + + + +
zma-miR160d-5p - - + + + + zma-miR169c-5p - - + + +
zma-miR160e - - + + + + zma-miR169r-5p - - + + +
zma-miR160f-5p - - + + + + zma-miR171d-3p - - - - -
zma-miR160g-5p - - + + + + zma-miR171e-3p - - - - -
zma-miR162-3p - - - - + - zma-miR171i-3p - - - - -
zma-miR164a-5p + - - + + zma-miR171j-3p - - - - -
zma-miR164b-5p + - - + + zma-miR171l-3p + - - - + +
zma-miR164c-5p + - - + + zma-miR171m-3p + - - - + +
zma-miR164d-5p + - - + + zma-miR172a - - + + - -
zma-miR164e-5p + + + - - zma-miR172b-3p - - + + - -
miRNA IM to SPM SPM to SM FM miRNA IM to SPM SPM to SM FM
zma-miR164f-5p + - - + + zma-miR172c-3p - - + + - -
zma-miR164g-5p + - - + + zma-miR172d-3p - - + + - -
zma-miR164h-5p + - - + + zma-miR172e - - + - -
zma-miR166a-3p - - + - + + zma-miR319a-3p - - + - -
zma-miR166b-3p - - + - + + zma-miR319b-3p - - + - -
zma-miR166c-3p - - + - + + zma-miR319c-3p - - + - -
zma-miR166d-3p - - + - + + zma-miR319d-3p - - + - -
zma-miR166e - - + - + + zma-miR390a-5p + + + +
zma-miR166f - - + - + + zma-miR390b-5p + + + +
zma-miR166g-3p - - + - + + zma-miR444a - - + - - -
zma-miR166h-3p - - + - + + zma-miR444b - - + - - -
zma-miR166i-3p - - + - + + zma-miR528a-5p + + + - + -
zma-miR166j-3p - - + - - zma-miR528b-5p + + + - + -
zma-miR166k-3p - - + - - zma-miR827-3p + - - -- - -
zma-miR166l-3p - - + - + +

表3

降解组测序检测到的玉米雌穗杂种优势相关miRNA的靶基因"

miRNA 靶基因
Target gene
分值
Score
校准范围
Alignment
range
切割位点
Cleavage
site
P
P-value
GO注释
Annotation for GO
zma-miR156 GRMZM2G414805_T02 1 1546-1565 1556 0.056054 SPL protein 11
zma-miR156 GRMZM2G414805_T03 1 1875-1894 1885 0.056054 SPL protein 11
zma-miR156 GRMZM2G414805_T04 1 1353-1372 1363 0.056054 SPL protein 11
zma-miR156 GRMZM2G414805_T05 1 2100-2119 2110 0.056054 SPL protein 11
zma-miR159 AC209015.3_FGT004 2.5 328-348 339 0.019582 non
zma-miR159 GRMZM2G028054_T02 3 1419-1439 1430 0.034972 transcription factor GAMYB
zma-miR159 GRMZM2G028054_T03 3 1360-1380 1371 0.034972 transcription factor GAMYB
zma-miR159 GRMZM2G075064_T01 2.5 851-871 862 0.019582 DNA binding
zma-miR159 GRMZM2G423833_T01 2 905-925 916 0.005424 DNA binding
zma-miR159 GRMZM2G139688_T01 3.5 1399-1419 1410 0.031147 DNA binding
zma-miR160 AC207656.3_FGT002 1 1350-1370 1361 0.037830 regulation of transcription
zma-miR160 GRMZM2G081406_T01 2 1499-1519 1510 0.005424 regulation of transcription
zma-miR160 GRMZM2G153233_T01 1 1652-1672 1663 0.037830 auxin response factor 2
zma-miR160 GRMZM2G159399_T01 1 1746-1766 1757 0.037830 regulation of transcription
zma-miR160 GRMZM2G390641_T01 1 1790-1810 1801 0.037830 regulation of transcription
zma-miR160 GRMZM2G390641_T02 1 1690-1710 1701 0.037830 regulation of transcription
zma-miR160 GRMZM5G808366_T01 1.5 1295-1315 1306 0.005915 auxin response factor 5
zma-miR160 GRMZM2G063522_T01 2 895-915 906 0.007152 regulation of transcription
zma-miR164 GRMZM2G139700_T01 2 737-757 748 0.026826 Nam2 protein
miRNA 靶基因
Target gene
分值
Score
校准范围
Alignment
range
切割位点
Cleavage
site
P
P-value
GO注释
Annotation for GO
zma-miR164 GRMZM2G393433_T01 2 854-874 865 0.026826 CUC2
zma-miR164 GRMZM2G393433_T02 2 814-834 825 0.026826 CUC2
zma-miR166 GRMZM2G003509_T01 3 736-756 747 0.068720 regulation of transcription
zma-miR166 GRMZM2G003509_T02 3 805-825 816 0.068720 regulation of transcription
zma-miR166 GRMZM2G042250_T01 3 628-648 639 0.068720 rolled leaf 2
zma-miR166 GRMZM2G042250_T02 3 714-734 725 0.068720 rolled leaf 2
zma-miR166 GRMZM2G042250_T03 3 628-648 639 0.068720 rolled leaf 2
zma-miR166 GRMZM2G042250_T04 3 628-648 639 0.068720 rolled leaf 2
zma-miR166 GRMZM2G109987_T01 3 1039-1059 1050 0.068720 transcription factor activity
zma-miR166 GRMZM2G109987_T02 3 1039-1059 1050 0.068720 transcription factor activity
zma-miR166 GRMZM2G109987_T03 3 1039-1059 1050 0.068720 transcription factor activity
zma-miR166 GRMZM2G109987_T04 3 1039-1059 1050 0.068720 transcription factor activity
zma-miR166 GRMZM2G109987_T05 3 774-794 785 0.068720 transcription factor activity
zma-miR166 GRMZM2G123644_T01 4 545-565 556 0.020551 DNA binding
zma-miR166 GRMZM2G178102_T01 3 1035-1055 1046 0.068720 transcription factor activity
zma-miR166 GRMZM2G336718_T01 3 105-124 115 0.013260 DNA binding
zma-miR167 GRMZM2G078274_T03 4 2531-2552 2542 0.034017 regulation of transcription
zma-miR169 GRMZM2G000686_T01 3.5 2120-2140 2131 0.057604 CCAAT-binding factor
zma-miR169 GRMZM2G000686_T02 3.5 1010-1030 1021 0.057604 CCAAT-binding factor
zma-miR169 GRMZM2G000686_T03 3.5 1006-1026 1017 0.057604 CCAAT-binding factor
zma-miR169 GRMZM2G000686_T04 3.5 962-982 973 0.057604 CCAAT-binding factor
zma-miR169 GRMZM2G000686_T05 3.5 934-954 945 0.057604 CCAAT-binding factor
zma-miR169 GRMZM2G000686_T06 3.5 890-910 901 0.057604 CCAAT-binding factor
zma-miR169 GRMZM2G000686_T08 3.5 1106-1126 1117 0.057604 CCAAT-binding factor
zma-miR169 GRMZM2G000686_T09 3.5 991-1011 1002 0.057604 CCAAT-binding factor
zma-miR169 GRMZM2G000686_T10 3.5 1300-1320 1311 0.057604 CCAAT-binding factor
zma-miR169 GRMZM2G091964_T01 4 1317-1338 1329 0.010329 nuclear transcription factor Y subunit A-1
zma-miR169 GRMZM2G091964_T02 4 1314-1335 1326 0.010329 nuclear transcription factor Y subunit A-1
zma-miR169 GRMZM2G091964_T03 4 1429-1450 1441 0.010329 nuclear transcription factor Y subunit A-1
zma-miR169 GRMZM2G165488_T01 3 1586-1606 1597 0.068720 transcription factor activity
zma-miR169 GRMZM2G165488_T02 3 2140-2160 2151 0.068720 transcription factor activity
zma-miR169 GRMZM2G165488_T03 3 1357-1377 1368 0.068720 transcription factor activity
zma-miR169 GRMZM5G829103_T02 3 1815-1835 1826 0.068720 NTF-Y subunit A-10
zma-miR169 GRMZM5G829103_T03 3 1321-1341 1332 0.068720 NTF-Y subunit A-10
zma-miR169 GRMZM5G829103_T04 3 1965-1985 1976 0.068720 NTF-Y subunit A-10
zma-miR169 GRMZM5G829103_T06 3 1288-1308 1299 0.068720 NTF-Y subunit A-10
zma-miR169 GRMZM5G829103_T07 3 1360-1380 1371 0.068720 NTF-Y subunit A-10
zma-miR171 GRMZM2G037792_T01 1 1322-1342 1333 0.056205 hypothetical protein
zma-miR171 GRMZM2G098800_T01 1 920-940 931 0.056205 hypothetical protein
zma-miR171 GRMZM2G098800_T02 1 1391-1411 1402 0.056205 hypothetical protein
zma-miR171 GRMZM2G110579_T01 1 1078-1098 1089 0.056205 putative uncharacterized protein
zma-miR171 GRMZM2G418899_T02 1 45-65 56 0.056205 non
zma-miR171 GRMZM5G825321_T01 1 507-527 518 0.056205 putative uncharacterized protein
zma-miR171 GRMZM2G418899_T01 1 45-65 56 0.056205 non
miRNA 靶基因
Target gene
分值
Score
校准范围
Alignment
range
切割位点
Cleavage
site
P
P-value
GO注释
Annotation for GO
zma-miR171 GRMZM5G825321_T02 1 1460-1480 1471 0.056205 putative uncharacterized protein
zma-miR172 GRMZM2G076602_T01 3 898-917 908 0.039256 putative uncharacterized protein
zma-miR172 GRMZM2G174784_T01 3 2242-2261 2252 0.039256 regulation of transcription
zma-miR172 GRMZM2G174784_T03 3 2367-2386 2377 0.039256 regulation of transcription
zma-miR172 GRMZM5G862109_T01 1 1559-1578 1569 0.050120 indeterminate spikelet1
zma-miR172 GRMZM5G862109_T02 1 1685-1704 1695 0.050120 indeterminate spikelet1
zma-miR172 GRMZM5G862109_T03 1 1671-1690 1681 0.050120 indeterminate spikelet1
zma-miR172 GRMZM2G174784_T02 3 2370-2389 2380 0.039256 regulation of transcription
zma-miR172 GRMZM2G700665_T02 1 1773-1792 1783 0.050120 AP2 transcription factor
zma-miR319 AC205574.3_FGT006 4 1456-1475 1466 0.023935 mutant cincinnata
zma-miR319 GRMZM2G015037_T01 4 1735-1754 1745 0.023935 glutamate 5-kinase activity
zma-miR319 GRMZM2G020805_T01 3 1429-1448 1439 0.008860 hypothetical protein
zma-miR319 GRMZM2G180568_T01 3.5 852-871 862 0.023453 transcribed locus
zma-miR319 GRMZM2G412073_T01 3.5 232-251 242 0.023453 hypothetical protein
zma-miR390 GRMZM2G084821_T02 4 1210-1230 1221 0.047298 hypothetical protein
zma-miR390 GRMZM2G084821_T04 4 611-631 622 0.047298 hypothetical protein
zma-miR390 GRMZM2G084821_T05 4 634-654 645 0.047298 hypothetical protein
zma-miR528 GRMZM2G107562_T01 2.5 570-589 580 0.004932 copper ion binding
zma-miR827 GRMZM2G044788_T01 2.5 245-265 256 0.001304 hypothetical protein

图2

新的玉米miRNA的前体茎环结构 miR-n7a和miR-n7b享有共同的pre-miRNA和成熟miRNA序列, 它们的特征被各自的pre-miRNA的基因组位置所区分, 蓝色代表已配对, 红色表示未配对, 绿色代表成熟的新的miRNA。"

图3

新miRNA靶基因功能分类 图中数字表示基因个数。"

表4

新的miRNA及其特性"

新的miRNA
Novel miRNA
序列
Sequence (5°-3°)
位置
Location
自由能指数
MFEI
IM to SPM SPM to SM FM
zma-miR-n1 AAAAAGAAACGGAGAGAGTAG 1:104607732:104607838: + 1.615137 D + -
zma-miR-n2 GTTTCTTTTTATTAGTCGTTG 4:21820530:21820628: - 1.300130 D - - -
zma-miR-n3 ATCCGACGCAAACGAACAAGG 10:1924407:1924548: - 1.027248 -- + +
zma-miR-n4 ATCCGACGCAAACGAACAAGG 4:215015818:215015940: - 0.953020 -- + +
zma-miR-n5 ACACAGTTGCTGTGCTCGCCG 9:149911503:149911831: + 0.947278 D + + + +
zma-miR-n6 AGGGCTTGTTCGTTTACGTCG 4:189032126:189032271: - 0.875367 ++ -- + +
zma-miR-n7a GCCTGAGGTCGGACAGTCCGC 1:52926508:52926666: - 0.874422 - D -
zma-miR-n7b GCCTGAGGTCGGACAGTCCGC 1:52934608:52934766: - 0.874422 - D -

"

图4

玉米雌穗杂种优势相关miRNA的表达趋势"

图5

miRNA对玉米雌穗发育的杂种优势的调控网络"

表6

测序获得的表达量数据"

NA H16-RPM H17-RPM H18-RPM H19-RPM H20-RPM H21-RPM H22-RPM H23-RPM H24-RPM
zma-miR1432-5p 0.199941767 0.177102302 0.2273795 0.6338742 0 5.0137835 0.11827416 0.1136377 0.61734739
zma-miR156a-5p 0.649810743 0.590341006 1.7053465 1.8439978 1.7981694 1.3624412 0.17741123 0.3409131 1.23469477
zma-miR156b-5p 3.548966364 1.889091218 2.1032607 29.56159 4.7201947 43.434625 2.48375726 1.4772903 1.32967129
zma-miR156c 0.649810743 0.590341006 1.7053465 1.8439978 1.7981694 1.3624412 0.11827416 0.3409131 1.23469477
zma-miR156d-5p 4.398718873 2.538466325 2.7853993 29.964964 5.4844167 43.707113 3.78477297 2.386392 1.42464781
zma-miR156e-5p 0.29991265 0.413238704 0.8526732 0.2304997 0.6293593 1.1444506 0.11827416 0.1704566 0.94976521
zma-miR156f-5p 0.599825301 0.472272805 1.6485016 1.8439978 1.7082609 1.3079435 0.11827416 0.3409131 1.09222999
zma-miR156g-5p 0.599825301 0.472272805 1.6485016 1.8439978 1.7082609 1.3079435 0.11827416 0.3409131 1.09222999
zma-miR156h-5p 0.29991265 0.413238704 0.8526732 0.2304997 0.6293593 1.1444506 0.11827416 0.1704566 0.94976521
zma-miR156i-5p 0.29991265 0.413238704 0.7958284 0.2304997 0.6293593 1.1444506 0.11827416 0.1704566 0.94976521
zma-miR156j-5p 7.947685237 5.549205454 19.384105 8.7589895 35.918434 17.60274 19.8700581 39.148192 7.36068037
zma-miR156k-5p 33.59021685 20.1306283 6.1392473 52.784437 8.8559844 13.133933 23.6548311 6.4773496 3.65659605
zma-miR156l-5p 0.29991265 0.413238704 0.7958284 0.2304997 0.6293593 1.1444506 0.11827416 0.1704566 0.90227695
zma-miR159a-3p 110.8177243 61.6906351 164.3954 69.207542 37.851466 166.05433 56.8898687 60.512082 759.384772
zma-miR159b-3p 110.7677389 61.6906351 164.3954 69.207542 37.851466 166.05433 56.8898687 60.512082 759.289796
zma-miR159c-3p 0.099970883 0.118068201 0.3410693 0 0.0449542 0.2724882 0 0.0568189 0.75981217
zma-miR159d-3p 0.099970883 0.118068201 0.3410693 0 0.0449542 0.2724882 0 0.0568189 0.75981217
zma-miR159f-3p 110.5677971 61.631601 164.3954 69.207542 37.851466 165.61835 56.8898687 60.512082 758.957378
zma-miR159j-3p 110.5178117 61.631601 164.3954 69.207542 37.851466 165.61835 56.8898687 60.512082 758.862402
zma-miR159k-3p 110.5178117 61.631601 164.3954 69.207542 37.851466 165.61835 56.8898687 60.512082 758.862402
zma-miR160a-5p 8.347568771 0.767443307 0.2842244 0.4609994 1.1688101 1.1989482 2.83857973 6.6478062 1.94701868
zma-miR160b-5p 8.347568771 0.767443307 0.2842244 0.4609994 1.1688101 1.1989482 2.83857973 6.6478062 1.94701868
zma-miR160c-5p 8.347568771 0.767443307 0.2842244 0.4609994 1.1688101 1.1989482 2.83857973 6.6478062 1.94701868
zma-miR160d-5p 8.597495979 0.826477408 0.2842244 0.6338742 1.1688101 1.3624412 3.07512804 6.6478062 1.94701868
NA H16-RPM H17-RPM H18-RPM H19-RPM H20-RPM H21-RPM H22-RPM H23-RPM H24-RPM
zma-miR160e 8.597495979 0.826477408 0.2842244 0.6338742 1.1688101 1.3624412 3.07512804 6.6478062 1.94701868
zma-miR160f-5p 8.597495979 0.826477408 0.2842244 0.6338742 1.1688101 1.3624412 3.07512804 6.6478062 1.94701868
zma-miR160g-5p 8.597495979 0.826477408 0.2842244 0.6338742 1.1688101 1.3624412 3.01599096 6.6478062 1.94701868
zma-miR162-3p 5.548384033 0.826477408 1.8190362 1.7863729 3.9559727 7.575173 2.1289348 1.5341091 4.3214317
zma-miR164a-5p 138.4596736 121.6692813 472.32413 101.36225 53.405632 262.07919 150.385589 62.61438 583.773185
zma-miR164b-5p 138.2597319 121.6692813 472.21044 101.247 53.315723 262.07919 150.326451 62.500742 583.725697
zma-miR164c-5p 138.2597319 121.6692813 472.21044 101.247 53.315723 262.07919 150.326451 62.500742 583.725697
zma-miR164d-5p 138.4596736 121.6692813 472.32413 101.36225 53.405632 262.07919 150.385589 62.61438 583.773185
zma-miR164e-5p 1.349606927 2.833636828 0.5116039 6.5116172 1.4834898 4.9592859 5.91370777 4.1477765 0.61734739
zma-miR164f-5p 138.4596736 121.6692813 472.32413 101.36225 53.360677 262.02469 150.385589 62.61438 583.725697
zma-miR164g-5p 138.2597319 121.6692813 472.21044 101.247 53.315723 262.07919 150.326451 62.500742 583.725697
zma-miR164h-5p 138.159761 121.3741108 471.86937 100.95888 53.135906 261.8067 150.267314 62.330286 583.013373
zma-miR166a-3p 8373.411244 2051.316927 5638.3302 6288.3206 2007.836 7944.231 3770.52094 2120.082 14058.0447
zma-miR166b-3p 8373.211302 2051.316927 5638.3302 6288.3206 2007.836 7944.3945 3770.4618 2120.082 14058.0447
zma-miR166c-3p 8373.161317 2051.316927 5638.3302 6288.3206 2007.836 7944.231 3770.40266 2120.082 14057.9972
zma-miR166d-3p 8373.211302 2051.316927 5638.3302 6288.3206 2007.836 7944.3945 3770.4618 2120.082 14058.0447
zma-miR166e 8373.261288 2051.316927 5638.3302 6288.3206 2007.8809 7944.7215 3770.4618 2120.082 14058.3296
zma-miR166f 8344.269731 2030.77306 5603.4843 6260.6606 1996.6424 7897.3631 3746.39301 2103.0932 13980.4489
zma-miR166g-3p 8344.219746 2030.832094 5603.5412 6260.6606 1996.5075 7897.3631 3746.39301 2102.9795 13980.4014
zma-miR166h-3p 8344.319717 2030.77306 5603.4843 6260.6606 1996.6424 7897.3631 3746.39301 2103.0932 13980.4489
zma-miR166i-3p 8372.011652 2051.021756 5636.2838 6287.1105 2007.7461 7940.4707 3769.33819 2119.8547 14050.874
zma-miR166j-3p 753.730476 307.8628345 265.18138 621.25438 424.81753 464.53794 401.185935 436.36882 312.187824
zma-miR166k-3p 754.4802576 307.7447663 264.954 621.94588 425.08725 465.19192 401.008524 436.59609 310.905641
zma-miR166l-3p 8375.410662 2055.626416 5639.1261 6288.8392 2008.7351 7947.1194 3772.82728 2121.6729 14058.0922
zma-miR166m-3p 8341.220619 2033.901867 5603.9959 6260.142 1995.5185 7898.889 3747.63489 2101.9568 13977.4571
zma-miR166n-3p 753.9304178 307.9218686 265.18138 621.312 425.0423 464.59244 401.245072 436.36882 312.235312
zma-miR167a-5p 6383.290867 3235.540984 1168.3329 4172.2755 864.29013 1672.2603 5075.91279 1339.9591 989.085488
zma-miR167b-5p 6383.390837 3235.304848 1168.3897 4172.3331 864.20022 1672.2603 5075.79451 1339.9591 989.085488
zma-miR167c-5p 6383.290867 3235.540984 1168.3329 4172.2755 864.29013 1672.2603 5075.91279 1339.9591 989.085488
zma-miR167d-5p 6383.290867 3235.540984 1168.3329 4172.2755 864.29013 1672.2603 5075.91279 1339.9591 989.085488
zma-miR167e-5p 6383.090925 3235.422916 1168.1055 4172.4484 864.29013 1672.1513 5074.84832 1340.1864 988.943023
zma-miR167f-5p 6383.14091 3235.422916 1168.1055 4172.4484 864.29013 1672.1513 5074.84832 1340.1864 988.943023
zma-miR167g-5p 6382.391129 3234.950643 1167.935 4171.8145 864.15527 1672.1513 5074.43436 1339.9591 988.943023
zma-miR167h-5p 6383.340852 3235.540984 1167.9918 4172.6789 864.20022 1672.5328 5075.02573 1340.5841 988.943023
zma-miR167i-5p 6383.390837 3235.600018 1167.9918 4172.6789 864.20022 1672.5328 5075.02573 1340.5841 988.943023
zma-miR167j-5p 6382.741027 3235.009677 1167.935 4171.8145 864.20022 1672.1513 5074.61177 1339.9591 988.990512
zma-miR168a-5p 5945.968237 5786.1093 7505.1162 13021.563 6575.9505 6315.2964 9509.95174 8001.7996 14166.9828
zma-miR168b-5p 5945.968237 5786.1093 7505.1162 13021.563 6575.9505 6315.2964 9509.95174 8001.7996 14166.9828
zma-miR169a-5p 0.699796184 0.944545609 3.9791418 2.2473723 1.3486271 1.9619153 0.35482247 1.6477468 3.37166649
zma-miR169b-5p 0.699796184 0.944545609 3.9791418 2.2473723 1.3486271 1.9619153 0.35482247 1.6477468 3.37166649
zma-miR169c-5p 0.699796184 0.590341006 2.5580197 2.1897474 0.9889932 0.9809577 0.35482247 1.4772903 2.32692476
NA H16-RPM H17-RPM H18-RPM H19-RPM H20-RPM H21-RPM H22-RPM H23-RPM H24-RPM
zma-miR169f-5p 4.248762548 2.06619352 0.2842244 10.141988 4.6302863 0.1089953 1.30101571 2.7273051 0.2374413
zma-miR169g 4.248762548 2.06619352 0.2842244 10.141988 4.6302863 0.1089953 1.30101571 2.7273051 0.2374413
zma-miR169h 4.248762548 2.06619352 0.2842244 10.141988 4.6302863 0.1089953 1.30101571 2.7273051 0.2374413
zma-miR169i-5p 4.248762548 2.06619352 0.2842244 10.141988 4.6302863 0.1089953 1.30101571 2.7273051 0.2374413
zma-miR169j-5p 4.248762548 2.06619352 0.2842244 10.141988 4.6302863 0.1089953 1.30101571 2.7273051 0.2374413
zma-miR169k-5p 4.248762548 2.06619352 0.2842244 10.141988 4.6302863 0.1089953 1.30101571 2.7273051 0.2374413
zma-miR169p-5p 4.29874799 2.00715942 0.2842244 10.199613 4.6302863 0.1089953 1.36015279 2.7273051 0.2374413
zma-miR169r-5p 0.699796184 0.590341006 2.5580197 2.1897474 0.9889932 0.9809577 0.35482247 1.4772903 2.32692476
zma-miR171b-3p 0.099970883 0.118068201 0.2842244 0.1152499 0.1348627 0.5994741 0.11827416 0 0.56985913
zma-miR171b-5p 0.099970883 0 0.6821386 0.2881247 0 1.2534459 0.17741123 0.1704566 0.56985913
zma-miR171d-3p 11.09676807 4.191421141 52.126757 11.582611 3.0119338 44.797066 8.51573919 2.8977617 30.4399749
zma-miR171d-5p 8.447539654 1.121647911 43.031576 6.8573668 1.0339474 51.990755 6.15025608 1.2500148 102.194736
zma-miR171e-3p 11.09676807 4.191421141 52.126757 11.582611 3.0119338 44.797066 8.51573919 2.8977617 30.4399749
zma-miR171e-5p 8.647481421 1.239716112 44.111629 6.9726166 0.9889932 52.099751 5.97284485 1.2500148 103.049525
zma-miR171f-3p 0.099970883 0.118068201 0.2842244 0.1152499 0.1348627 0.5994741 0.11827416 0 0.56985913
zma-miR171h-3p 0.699796184 0.059034101 0.3979142 0.5186244 0.0899085 0.3269859 0.47309662 0.2272754 0.09497652
zma-miR171h-5p 1.14966516 0.767443307 0.2842244 0.1152499 0.0449542 0.1089953 0.59137078 0.4545509 0.09497652
zma-miR171i-3p 11.09676807 4.13238704 52.013068 11.582611 3.056888 44.524578 8.51573919 2.8977617 30.2500219
zma-miR171j-3p 11.09676807 4.13238704 52.126757 11.582611 3.056888 44.797066 8.51573919 2.8977617 30.4399749
zma-miR171k-3p 0.699796184 0.059034101 0.3979142 0.5186244 0.0899085 0.3269859 0.47309662 0.2272754 0.09497652
zma-miR171k-5p 1.14966516 0.767443307 0.2842244 0.1152499 0.0449542 0.1089953 0.59137078 0.4545509 0.09497652
zma-miR171l-3p 1.549548694 1.121647911 5.4571087 2.1321224 0.6743135 9.7550789 1.83324941 0.2840943 12.7743421
zma-miR171m-3p 1.549548694 1.121647911 5.4571087 2.1321224 0.6743135 9.7550789 1.83324941 0.2840943 12.7743421
zma-miR172a 30.49111946 12.86943393 4.2633662 155.64494 15.913799 6.2672294 24.0096535 24.148014 0.14246478
zma-miR172b-3p 30.5411049 12.86943393 4.2633662 155.64494 15.913799 6.2672294 24.0096535 24.148014 0.14246478
zma-miR172c-3p 30.5411049 12.86943393 4.2633662 155.64494 15.913799 6.2672294 24.0096535 24.148014 0.14246478
zma-miR172d-3p 30.5411049 12.86943393 4.2633662 155.64494 15.913799 6.2672294 24.0096535 24.148014 0.14246478
zma-miR172e 4.348733431 6.375682862 1.8758811 6.684492 16.273433 0.5994741 3.37081343 16.079736 0.37990608
zma-miR319a-3p 29.89129416 9.327387891 1.3074323 28.178591 23.690882 1.1444506 9.69848074 23.523007 0.37990608
zma-miR319b-3p 30.14122137 9.386421992 1.5916567 28.87009 24.185379 1.4169388 9.81675489 23.977557 0.52237086
zma-miR319c-3p 29.84130872 9.327387891 1.3074323 28.178591 23.690882 1.1444506 9.69848074 23.523007 0.37990608
zma-miR319d-3p 30.14122137 9.386421992 1.5916567 28.87009 24.185379 1.4169388 9.81675489 23.977557 0.52237086
zma-miR390a-5p 21.64369627 11.21647911 38.142916 29.33109 14.789943 50.355826 30.455595 15.341091 136.908655
zma-miR390b-5p 21.64369627 11.21647911 38.142916 29.33109 14.789943 50.355826 30.455595 15.341091 136.908655
zma-miR395b-5p 2.049403111 3.483011934 0 0 0 0 2.83857973 1.3636526 0
zma-miR395e-5p 0.899737951 4.07335294 0 0.0576249 0 0 2.36548311 1.3636526 0
zma-miR395h-5p 0.899737951 4.07335294 0 0.0576249 0 0 2.36548311 1.3636526 0
zma-miR395i-5p 2.049403111 3.483011934 0 0 0 0 2.83857973 1.3636526 0
zma-miR395j-5p 0.899737951 4.07335294 0 0.0576249 0 0 2.36548311 1.3636526 0
zma-miR395p-5p 0.899737951 4.07335294 0 0.0576249 0 0 2.36548311 1.3636526 0
zma-miR396a-5p 0.249927209 0.472272805 0.2842244 0.9796238 0.2247712 0.1634929 0.17741123 0.4545509 0.04748826
NA H16-RPM H17-RPM H18-RPM H19-RPM H20-RPM H21-RPM H22-RPM H23-RPM H24-RPM
zma-miR396b-5p 0.249927209 0.472272805 0.2842244 0.9796238 0.2247712 0.1634929 0.17741123 0.4545509 0.04748826
zma-miR396c 0.049985442 0 3.6949174 0.4609994 0 3.5968447 0.05913708 0.1136377 5.69859125
zma-miR396d 0.049985442 0 3.6949174 0.4609994 0 3.5968447 0.05913708 0.1136377 5.69859125
zma-miR396e-5p 0.199941767 0.472272805 0.2842244 0.864374 0.1798169 0.1634929 0.17741123 0.4545509 0.04748826
zma-miR396f-5p 0.199941767 0.472272805 0.2842244 0.864374 0.1798169 0.1634929 0.17741123 0.4545509 0.04748826
zma-miR397a-5p 0.099970883 0.236136402 0.4547591 0.0576249 0 0.4904788 0.29568539 0.1704566 0
zma-miR397b-5p 0.099970883 0.236136402 0.5684488 0.0576249 0 0.4904788 0.29568539 0.2840943 0.04748826
zma-miR398a-3p 0.149956325 0.059034101 4.831815 0.2304997 0.0449542 1.85292 0.05913708 0.0568189 0.52237086
zma-miR398b-3p 0.149956325 0.059034101 4.831815 0.2304997 0.0449542 1.9074177 0.05913708 0.0568189 0.52237086
zma-miR408b-5p 0.549839859 0.531306905 0.966363 1.0372488 0.2247712 1.5804318 1.30101571 0.795464 1.13971825
zma-miR444a 3.249053713 1.889091218 7.1056103 3.6879956 0.5844051 4.0873235 1.77411233 1.2500148 4.7963143
zma-miR444b 3.299039155 2.184261721 7.3329899 3.8608704 0.8990847 4.1418212 1.83324941 1.4772903 5.12873213
zma-miR528a-5p 512.3007924 282.6552736 25.693887 1062.7766 148.75357 220.71547 1562.28332 207.38883 113.876849
zma-miR528b-5p 512.4507487 282.89141 25.750732 1062.8918 148.75357 220.76997 1562.63814 207.55928 113.876849
zma-miR529-5p 45.18683933 5.07693265 0.3410693 27.371842 0.5844051 0.5449765 25.0149839 0.6818263 0.2374413
zma-miR827-3p 15.29554517 138.3759317 22.283194 14.809607 27.107404 31.990119 31.3426512 58.296147 13.9140603
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